基因枪介导真核表达质粒转染体外培养的COS-7细胞系的实验研究

目的：建立基因枪子弹制备及转染体外培养COS-7细胞系的方法。方法：以亚精氨、氯化钙沉淀法制备子弹（DNA＋金颗粒）,利用原子力显微镜观察子弹制备情况;采用基因枪方法分别将真核表达质粒pVax-Dsred-IRES-EGFP转染对照组和实验组COS-7细胞,转染后24h,利用激光扫描共聚焦显微镜观察细胞中红、绿荧光蛋白的表达。结果：制备了基因枪子弹,DNA紧密包裹在金颗粒周围;基因枪介导的pVax-Dsred-IRES-EGFP被转染入体外培养的COS-7细胞,转染后24h可检测到红、绿荧光,而对照组则没有荧光蛋白的表达。结论：国产新芝SJ-500型基因枪能够有效介导外源基因转移,基因枪转染的COS-7细胞能够有效表达报告基因。

Empirical Study of Gene Gum-Mediated Eukaryotic Expression Vector Transfection of COS-7 Cell Lines/n Vitro

Objective： To establish a preparation method of cartridge for gene gun and to observe the gene gun-mediated gene transfectiono to COS-7 cell lines in vitro. Methods： The cartridge（microcarrier＋DNA） was prepared by precipitation method of spermidine and calcium chloride. Using atomic force microscope to observe the detail of the cartridge. The recombinant eukaryotie expression vector pVax-Dsred-IRES-EGFP was tansfected into COS-7 cell lines by gene gun. The. expression of report gene was observed through laser scan confocal microscopy. Results： The cartridge of gene gun was successfully prepared, DNA-coated gold mierocarriers was achieved. Red and green fluorescent protein could be detected in the transfected COS-7 cell lines after 24 hours, when no fluorescent protein was detected in control group. Conclusion： SJ-500 gene gun-mediated gene transfer is effective and practicable. The report gene can be expressed through gene gun mediated transfection to cell lines in vitro.